细胞内非编码RNA双成像分析新方法的研究及应用

Non-coding RNAs play key roles at all levels in gene expression, immune system, and tumorigenesis. Therefore, the sensitive detection of non-coding RNA, especially at cell or molecule level, has become a focus and hotspot in recent year's research. With the fast development of imaging technology, nanomaterials owned excellent properties have been widely applied in the fields of imaging analysis. However, some shortcomings of imaging technology and nanomaterials, including low stability and sensitivity of probes, the “false positive” results caused by detection of a single target, light bleaching and cell photo damage during in imaging analysis, still exist in the present analytical methods. To address these problems, on basis of the recent established imaging methods of mRNA by our group, this project aims at the development of new optical probes owning bifunctional imaging properties, providing insight on fundamental understanding of the signal generation mechanism and transformation mechanism. The novel probes can be further combined with dark-field and two-photon microscope imaging technique, for establishing a dual-imaging analysis strategy for intracellular non-coding RNA. Besides, we plan to extend the application scope of this method, study the intracellular multiple analytes simultaneous and dynamical imaging analysis in real-time, meanwhile imaging at molecule level, thereby provide powerful techniques for biochemical analysis.

非编码RNA (non-coding RNA)在基因表达、机体免疫及肿瘤发生等各个层面均发挥重要调控功能，因此non-coding RNA 的高灵敏分析，特别是在细胞和分子层面上的成像分析，已成为人们研究的重点与热点。高速发展的成像技术和性能独特的纳米材料成为上述研究中不可或缺的工具，但目前的研究方法仍面临以下诸多问题：探针的稳定性和灵敏度差，单一分析的假阳性，光漂白及成像过程中细胞的光损伤等。本项目针对这些问题，以我们前期所建立的细胞内mRNA成像分析研究为基础，旨在进一步研发新型的具有双成像功能的光学纳米检测探针，深入研究双成像信号的产生机制和转化机制。在此基础上，将探针与暗场成像和双光子成像技术相结合，建立non-coding RNA原位双成像分析新方法，并进一步扩大这种方法的应用范围，研究活细胞内多组分的同时检测及分子层面上的成像分析，为开展生化分析提供强而有力的技术支持。

非编码RNA（non-coding RNA）在基因表达、细胞调控、肿瘤发生及治疗等各个层面中都发挥重要的作用，因此实现此类RNA的成像分析对于探究肿瘤微环境的特征构成、疾病发生的分子基础及病理演化规律等具有极其重要的意义。然而RNA的同源性及生理微环境的复杂性极大的限制了对此类分子的细胞内原位成像分析。针对上述问题，在本项目的支持下，系统的开展了两个方面的工作：首先将多功能贵金属纳米材料及二维片层纳米材料与成像技术相结合，实现了不同肿瘤细胞内miRNA（代表性的非编码RNA）的高灵敏可视化分析；在此基础上，将多重成像技术相结合，利用非编码RNA及肿瘤微环境作为触控因子，实现了对肿瘤细胞及组织的靶向诊疗。综上所述，此类成像分析方法及多模式成像诊疗技术的构建，为恶性肿瘤病理学研究及诊疗提供理论思路和强而有力的技术支持。